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Advances in informatics, robotics, and imaging techniques make it possible to use state-of-the-art digital reconstruction technologies for high-throughput plant phenotyping (HTPP) affected by stress factors, as well as for the ontology of their structural and functional traits. Digital imaging of structural and functional features of the aboveground part of plants is non-destructive and plants can be monitored throughout their entire life cycle. In the experiment with tomato plants (Solanum lycopersicum L.; cv. Gruzanski zlatni) grown in controlled environmental conditions and affected by gradual soil dehydration, we evaluated phenotypic traits and phenotypic plasticity by the PlantScreenTM platform using digital imaging of plant optical signals. In this study, 25 different morpho-physiological traits of the plant were evaluated during the precise control and monitoring of the water content in the soil. Different levels of plant water supply induced statistically significant differences in the formation of individual phenotypic traits. Several plant traits have been identified that are characterized by low variability in both well-hydrated and water-stressed conditions, as well as traits with high phenotypic plasticity. Geometric traits (especially Isotop, Round-2top, and Compside) showed a relatively low level of drought-induced phenotypic plasticity. However, functional and chemometric characteristics (ΔF/F′m, Rfd, Water-1, and ARI-1) showed the potential to exhibit rapid plasticity in water-stressed conditions. Our results confirmed that a high-throughput phenotyping methodology coupled with advanced statistical analysis tools can be successfully applied to characterize crop stress responses and identify traits associated with crop stress tolerance.

Glucose is the primary source of carbon and energy. Cells possess a sophisticated mechanism for sensing glucose and responding to it appropriately. However, hyperglycemia represents the predominant risk factor for the development of diabetes. On the other hand, nutrient starvation, including glucose, may lead to the development of malnutrition. Noteworthy, glucose restriction has been associated with significantly delayed aging. Thus, controlled glucose intake is vital for healthy living. The presented study shows the effect of the oyster mushroom Pleurotus ostreatus ethanolic extract on fission yeast Schizosaccharomyces pombe undergoing glucose-derived stress. Fruiting bodies of P. ostreatus were used for ethanolic extraction and its anti-hyper- and hypo-glycemic stress activity was investigated. Compared were cells treated with ethanolic extract of P. ostreatus (1 or 2%) with untreated control. Different glucose concentrations in growth media were used to analyze its impact on growth intensity, generation time (gt), metabolic activity, ROS generation, MDA content, and cell death and apoptosis of S. pombe cells. Glucose deprivation or its over-supplementation resulted in growth retardation and prolonged cell doubling time (gt) which was significantly adjusted by the oyster mushroom extract. Interestingly, oyster mushroom extract reduced excessive ROS and formed MDA that were produced as a consequence of glycemic stress. Unexpectedly, the extract was able to increase the metabolic activity of cells supplemented with high glucose content and thereby protected them from death. Taken together, our study suggests that growth media supplementation with the ethanolic extract of P. ostreatus protects S. pombe cells from glucose-mediated stress through alleviation of oxidative stress and enhancement of metabolic activity. [ABSTRACT FROM AUTHOR]

The application of biological active preparations (BAPs) and remote-sensing control in the management of agronomic intervention are an important part of successful crop cultivation. The effects of foliar application of two BAPs (containing amino acids or Abiestins®) on yield and yield-forming, as well eco-physiological traits calculated from infrared thermographs data (crop water stress index, CWSI and index of stomatal conductance, Ig) of three hybrids of sunflower were studied in field poly-factorial experiments, realised during two years (2012 and 2013). The results showed that the application of selected BAPs has contributed to an increase of the sunflower yield, in particular through an increase in the weight of thousand seeds (rp = 0.761, P < 0.001). Similarly, oil content in achenes was significantly higher in treatments with BAPs, mainly with preparation containing free amino acids. The study describes the quantitative relationship between yield and quality of sunflower production (rp = −0.41, P < 0.01). Selected hybrids of sunflower in two growth stages showed the significant differences in CWSI and Ig (both at P < 0.01), respectively. An analysis of negative linear relation between the yield of achenes and CWSI (rp = −0.654, P < 0.001) confirmed that higher value of plant stress resulted in a smaller yield and vice-versa. The opposite trend was observed between yield and Ig index (rp = 0.576, P < 0.001). The data obtained from IR thermography can be used for monitoring the physiological health of sunflower plants, as well in potential prediction and control of yield.

Cadmium (Cd) a highly toxic environmental pollutant, that does not have any physiological function in the organism, represents a great concern for human health as it can be easily transported from its environmental sources to the food chain. Food, water, and air are the major sources of Cd exposure to the population. Cd-mediated impairments of the basic cellular properties largely depend on its ability to enhance the formation of reactive oxygen species (ROS) and thus triggers oxidative stress to the cell. With the use of fission yeast Schizosaccharomyces pombe (S. pombe) as the model organism, we have analyzed the impact of Cd on the cell growth intensity, as it represents the fundamental feature of all living organisms. Cells were incubated with different Cd concentrations for 3, 6, and 9 hours to investigate the effect of Cd on cell growth in a time and dose-dependent manner. Further possible Cd-derived alterations, as the peroxidation of membrane lipids or the functional impairment of the enzymatic antioxidant protection mechanisms, were investigated by determination of the MDA content and via catalase (CAT) activity detection. Moreover, ascorbic acid (AsA) pre-treatment was subjected to investigate the assumed positive effect of AsA against Cd toxicity. We show here on one hand that cells suffer under the influence of Cd, but on the other hand, they substantially profit from AsA supplementation. Because S. pombe is known to shares many molecular, and biochemical similarities with higher organisms, the effect of AsA in cadmium toxicity elimination might be expected to a similar extent also in other cell types. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Central European Agriculture is the property of Journal of Central European Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Central European Agriculture is the property of Journal of Central European Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Central European Agriculture is the property of Journal of Central European Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Listy Cukrovarnicke a Reparske is the property of VUC Praha a.s. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Listy Cukrovarnicke a Reparske is the property of VUC Praha a.s. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Listy Cukrovarnicke a Reparske is the property of VUC Praha a.s. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Doxorubicin is one of the most potent anti-tumor drugs with a broad spectrum of use. To reduce its toxic effect and improve its pharmacokinetics, we conjugated it to an HPMA copolymer carrier that enhances its passive accumulation within solid tumors via the EPR effect and decreases its cytotoxicity to normal, noncancer cells. In this study, we compared the antiproliferative, pro-survival, and death signals triggered in EL-4 cancer cells exposed to free doxorubicin and doxorubicin conjugated to a HPMA copolymer carrier via either enzymatically (PK1) or hydrolytically (HYD) degradable bonds. We have previously shown that the intracellular distribution of free doxorubicin, HYD, and PK1 is markedly different. Here, we demonstrated that these three agents greatly differ also in the antiproliferative effect and cell death signals they trigger. JNK phosphorylation sharply increased in cells treated with HYD, while treatment with free doxorubicin moderately decreased and treatment with PK1 even strongly decreased it. On the other hand, treatment with free doxorubicin greatly increased p38 phosphorylation, while PK1 and HYD increased it slightly. PK1 also significantly increased ERK phosphorylation, while both the free doxorubicin and HYD conjugate slightly decreased it. Long-term inhibition of JNK significantly increased both proliferation and viability of EL-4 cells treated with free doxorubicin, showing that the JNK signaling pathway could be critical for mediating cell death in EL-4 cells exposed to free doxorubicin. Both activation of caspase 3 and decreased binding activity of the p50 subunit of NFkappaB were observed in cells treated with free doxorubicin and HYD, while no such effects were seen in cells incubated with PK1. Analysis of the expression of genes involved in apoptosis and regulation of the cell cycle demonstrated that free doxorubicin and HYD have very similar mechanisms of action, while PK1 has very different characteristics.

An N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer carrier containing doxorubicin and human immunoglobulin as an actively/passively targeting moiety was used in four patients with generalized breast cancer resistant to standard cytotoxic chemotherapy. The dose and time schedule were deduced from a Phase I clinical trial in which doxorubicin bound to HPMA copolymer carrier (PK1) was tested. It was confirmed that the Dox-HPMA-HuIg conjugate is stable and doxorubicin remains in the peripheral blood with a small amount also in the urine, mostly in its polymer-bound form. More than 116 biochemical, immunological and hematological parameters were determined for blood samples taken from patients 24 h, 48 h, 72 h and 1 to 11 weeks after treatment. Depending on the patient, some parameters decreased permanently or temporarily to the normal level (CRP, C3, CA 72-4, beta(2)-microglobulin, ferritin, CEA, CA 125, CD4, CD8, CE19, CD16(+)56(+), leu, ery) and some moved markedly towards physiological values (AST, ALT, ALP, GMT, CA 15-3, NSE, AFP). While the number of peripheral blood reticulocytes was significantly decreased after treatment with the classical free drug, their number was not affected or was even elevated after treatment with Dox-HPMA-HuIg. Increased absolute numbers of CD16(+)56(+) and CD4(+) cells in the peripheral blood and activation of NK and LAK cells in all patients support data obtained in experimental animals, pointing to a dual, i.e. cytostatic and immunomobilizing character of Dox-HPMA conjugates containing a targeting immunoglobulin moiety.

This study proposes a strategy to generate new anticancer therapy using hydrogel-based drug delivery systems to improve drug bioavailability and increase the therapeutic efficacy. We have synthesized biodegradable hydrogels based on N-(2-hydroxypropyl)methacrylamide (HPMA) with prolonged drug release. Pharmacokinetic data from in vitro studies showed that the in vitro release of hydrophilic drugs (doxorubicin, vinblastine) from HPMA-hydrogels is affected mainly by drug diffusion and only partially by hydrogel degradation. The release of hydrophobic drugs (cyclosporine A, CsA) actually copies the process of degradation and therefore it is slower. Hydrogels with degradation time of 50 h released the doxorubicin over a period of at least 96 h after s.c. implantation. Drug concentration at pharmacologically active levels was maintained in the bloodstream over a period of at least 4 days, ranging between 0.1 and 1 microg/ml. The therapeutic potential of HPMA-hydrogels in vivo was studied in Bcl1 leukemia. HPMA-hydrogels containing DOX were significantly more effective in inhibition of Bcl1 leukemia in comparison with free DOX or non-targeted polymeric drug (PK1). The efficacy of therapeutic combination using unspecific, hydrogel-based therapy with specific, antibody-targeted therapy at late stages of Bcl1 leukemia was also tested. In contrast to application of DOX alone, a cocktail of DOX with CsA as a blocker of P-glycoprotein (Pgp) incorporated into HPMA-hydrogel blocked the proliferation of Pgp-overexpressing multidrug resistant cell lines in vitro by induction of apoptosis.

The aim of this study was to compare the properties and antitumor potential of a novel type of antibody-targeted N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-bound doxorubicin conjugates with star structure with those of previously described classic antibody-targeted or lectin-targeted HPMA copolymer-bound doxorubicin conjugates. Classic antibody-targeted conjugates were prepared by aminolytic reaction of the multivalent HPMA copolymer containing side-chains ending in 4-nitrophenyl ester (ONp) reactive groups with primary NH(2) groups of the antibodies. The star structure of antibody-targeted conjugates was prepared using semitelechelic HPMA copolymer chains containing only one reactive N-hydroxysuccinimide group at the end of the backbone chain. In both types of conjugates, B1 monoclonal antibody (mAb) was used as a targeting moiety. B1 mAb recognizes the idiotype of surface IgM on BCL1 cells. The star structure of the targeted conjugate had a narrower molecular mass distribution than the classic structure. The peak in the star structure was around 300-350 kDa, while the classic structure conjugate had a peak around 1300 kDa. Doxorubicin was bound to the HPMA copolymer via Gly-Phe(D,L)-Leu-Gly spacer to ensure the controlled intracellular delivery. The release of doxorubicin from polymer conjugates incubated in the presence of cathepsin B was almost twice faster from the star structure of targeted conjugate than from the classic one. The star structure of the targeted conjugate showed a lower binding activity to BCL1 cells in vitro, but the cytostatic activity measured by [(3)H]thymidine incorporation was three times higher than that seen with the classic conjugate. Cytostatic activity of nontargeted and anti-Thy 1.2 mAb (irrelevant mAb) modified HPMA copolymer-bound doxorubicin was more than hundred times lower as compared to the star structure of B1 mAb targeted conjugate. In vivo, both types of conjugates targeted with B1 mAb bound to BCL1 cells in the spleen with approximately the same intensity. The classic structure of the targeted conjugate bound to BCL1 cells in the blood with a slightly higher intensity than the star structure. Both types of targeted conjugates had a much stronger antitumor effect than nontargeted HPMA copolymer-bound doxorubicin and free doxorubicin. The star structure of targeted conjugate had a remarkably higher antitumor effect than the classic structure: a single intravenous dose of 100 microg of doxorubicin given on day 11 completely cured five out of nine experimental animals whereas the classic structure of targeted conjugate given in the same schedule only prolonged the survival of experimental mice to 138% of control mice. These results show that the star structure of antibody-targeted HPMA copolymer-bound doxorubicin is a suitable conjugate for targeted drug delivery with better characterization, higher cytostatic activity in vitro, and stronger antitumor potential in vivo than classic conjugates.